Selection of reference genes for normalization of cranberry (Vaccinium macrocarpon Ait.) gene expression under different experimental conditions


Autoři: Chen Li aff001;  Jian Xu aff001;  Yu Deng aff002;  Haiyue Sun aff001;  Yadong Li aff001
Působiště autorů: Engineering Center of Genetic Breeding and Innovative Utilization of Small Fruits of Jilin Province, College of Horticulture, Jilin Agricultural University, Changchun, China aff001;  College of Life Sciences, Jilin Agricultural University, Changchun, China aff002
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224798

Souhrn

Real-time fluorescent quantitative PCR (qRT-PCR) is often chosen as an effective experimental method for analyzing gene expression. However, an appropriate reference gene as a standard is needed to obtain accurate gene expression data. To date, no internal reference genes have been reported for research on cranberries. Expanding the selection of internal reference genes for cranberry will enable reliable gene expression analysis, and, at the same time, can also lay a solid foundation for revealing the biological characteristics of cranberry. Here, we selected ten candidate reference gene families and used three statistical software tools—geNorm, NormFinder and BestKeeper—to evaluate their expression stability under the influence of different experimental factors. The results showed that protein phosphatase 2A regulatory subunit (PP2A) or RNA helicase-like 8 (RH 8) was the best choice for an internal reference gene when analyzing different cranberry cultivars. In two sample sets comprising different cranberry organs and three abiotic stress treatments, sand family protein (SAND) was the best choice as a reference gene. In this study, we screened genes that are stably expressed under the influence of various experimental factors by qRT-PCR. Our results will guide future studies involving gene expression analysis of cranberry.

Klíčová slova:

Fruits – Gene expression – Genetic screens – Leaves – Library screening – Plant resistance to abiotic stress – Ribosomal RNA – RNA extraction


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